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首页> 外文期刊>Folia biologica >New Luminescence-Based Approach to Measurement of Luciferase Gene Expression Reporter Activity and Adenosine Triphosphate-Based Determination of Cell Viability
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New Luminescence-Based Approach to Measurement of Luciferase Gene Expression Reporter Activity and Adenosine Triphosphate-Based Determination of Cell Viability

机译：基于荧光的荧光素酶基因表达报告基因活性测定新方法和基于三磷酸腺苷的细胞活力测定

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Renal ischaemia/reperfusion (I/R) injuryand hypertension represent major alloantigen-independent risk factors contributing to the developmentof chronic allograft nephropathy. In a model of accelerated major histocompatibility complex-independent renal injury, we evaluated the effect of leflunomide derivate – FK778 – on the progression ofaccelerated nephropathy. Thirty-six uninephrectomized hypertensive transgenic (m-REN-2)-27 ratsreceived a clip on renal pedicle for 45 minutes. Animals were treated with FK778 3 mg/kg/day (I/R3 mg, N = 12), 10 mg/kg/day (I/R 10 mg, N = 12) orplacebo (N = 12) via gavage for 16 weeks. Eighteenanimals were sham-operated and treated with FK7783 mg/kg/day (sham 3 mg, N = 6), 10 mg/kg/day (sham10 mg, N = 6) or were untreated (sham, N = 6). Proteinuria and blood pressure were evaluated throughout and the kidneys were harvested for morphological and immunohistochemical analysis at the end ofthe experiment. At week 16, rats with I/R injury andFK778 treatment had lower proteinuria comparedwith placebo-treated rats (I/R 3 mg: 48.42 ± 26.16,I/R 10 mg 27.28 ± 21.86 vs. Placebo: 70.13 ± 50.19mg/day, P < 0.05). The untreated sham group exhibited lower proteinuria compared with FK778-treated sham groups (Sham 3 mg: 24.23 ± 10.89; Sham10 mg: 17.37 ± 4.13; Sham: 14.23 ± 1.18) There wasno difference in glomerulosclerosis and interstitialfibrosis among the treated groups. In the untreatedanimals the rate of interstitial fibrosis decline reachedstatistical significance (Placebo vs. Sham: 1.125 ±0.641 % vs. 0.250 ± 0.500 %, P < 0.05). There washigher CD5+ leukocyte infiltration in the placebotreated group. FK778-treated rats displayed amelioration of some changes induced by the I/R injury.Our observation also suggests potential nephrotoxicity of FK778.
机译：肾缺血/再灌注（I / R）损伤和高血压代表了主要的同种异体抗原独立危险因素，导致慢性同种异体肾病的发展。在一个独立于主要组织相容性复合体的加速严重肾脏损伤模型中，我们评估了来氟米特衍生物FK778对加速性肾病进展的影响。 36只未切除肾小管的高血压转基因（m-REN-2）-27大鼠在肾蒂上夹了45分钟。通过管饲法以3 mg / kg /天（I / R3 mg，N = 12），10 mg / kg /天（I / R 10 mg，N = 12）或安慰剂（N = 12）FK778治疗动物16周。假手术的动物有18只，分别接受FK7783 mg / kg /天（假3 mg，N = 6），10 mg / kg /天（假10 mg，N = 6）或未经治疗（假，N = 6）。在整个实验过程中对蛋白尿和血压进行了评估，并在实验结束时收集肾脏进行形态学和免疫组化分析。与安慰剂治疗组相比，患有I / R损伤和FK778治疗的大鼠的蛋白尿水平较低（I / R 3 mg：48.42±26.16，I / R 10 mg 27.28±21.86，而安慰剂：70.13±50.19mg /天， P <0.05）。与FK778治疗的假手术组相比，未治疗的假手术组表现出较低的蛋白尿（假手术3 mg：24.23±10.89; Sham10 mg：17.37±4.13; Sham：14.23±1.18）在治疗组之间肾小球硬化和间质纤维化没有差异。在未经处理的动物中，间质纤维化的下降率达到统计学显着性（安慰剂与假手术：1.125±0.641％与0.250±0.500％，P <0.05）。安慰剂治疗组的CD5 +白细胞浸润较高。用FK778治疗的大鼠表现出I / R损伤引起的某些变化的改善，我们的观察结果还表明FK778具有潜在的肾毒性。

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